Study On Original Identification And Antitumor Activity Of Traditional Chinese Medicine Rhizoma Amorphophalli

Objective: Based on the original chaotic problem of Rhizoma Amorphophalli for clinical applications,this research establishes its traditional appraisal methods and molecular marker DNA barcode identification method,clarifies the medicinal material base of the original to provide a scientific basis for reasonable clinical medication safety.Meanwhile,this research selects Rhizoma Amorphophalli area of antitumor activity and sensitive tumor cells to provide the theory basis for basic research of Rhizoma Amorphophalli antitumor active ingredient of material,and to lay a foundation for the further research of Rhizoma Amorphophalli antitumor.Methods: This paper adopts the literature review method by reading ancient herbs records on the Rhizoma Amorphophalli,conducting basis of the original research,and collecting Rhizoma Amorphophalli herbal medicine standards and processing standards in different areas.It collected a total of 21 samples of various samples through field research.The identification of the basal roots of the snake basins was carried out by using the identification of original plant traits,microscopic identification,identification of TLC,determination of content and establishment of specific primers polymerase chain reaction DNA bar code molecular identification technology to comprehensively improve its quality standard,and enhance quality control.By the method of CCK 8 in vitro cultured human ovarian cancer,breast cancer,liver cancer and stomach cancer cell to Rhizoma Amorphophalli antitumor activity in vitro studies and solvent to the system of Rhizoma Amorphophalli different polar parts activity screening,and through the fluorescence inverted microscope to observe the cell apoptosis cells morphological changes and the detection further studies the Rhizoma Amorphophalli active site,tumor value-added activities.Results:1.The collected 21 Rhizoma Amorphophalli samples were identified by literature and identified by the traditional method to find out the difference of interspecific traits,and determined the She liu gu originally base of medicinal plant of Araceae Amorphophallus konjac K.Koch and Amorphophallus sinensis Belval,after microscopicidentification,thin layer identification research prove that the sample number 1,3,4,6,8,9,11,12,13,14,15,N,S,X,Y,G,J for the Amorphophallus konjac K.Koch,sample number2,5,7 were falsify species,sample number 10 traits similar to Rhizoma Amorphophalli characters,but no TLC behavior,not identified;Through DNA molecular identification of bar code technology,the Rhizoma Amorphophalli medicinal base the original Amorphophallus sinensis Belval designed specific primer polymerase chain reaction,the establishment of a DNA barcode molecular identification method can quickly detect Rhizoma Amorphophalli medicinal plants Amorphophallus sinensis Belval.2.In vitro antitumor activity studies showed that Rhizoma Amorphophalli had a good inhibitory effect on human ovarian cancer cells,human breast cancer,human hepatocellular carcinoma cells and human gastric cancer cells,and showed a good dose-time effect relationship.The inhibitory rate and IC50 value of the four kinds of tumor cells cultured in vitro showed that the ethyl acetate site had high inhibitory activity,and the IC50 value of human ovarian cancer cells was the lowest,0.766mg/ml,valley anti-tumor activity is mainly concentrated in the ethyl acetate site;After observing the morphology of the cells with different concentrations of ethyl acetate extract on the above four kinds of cells,the morphology of the cells was observed by fluorescence uptake microscope.The cell morphology was abnormal,the volume was reduced,the original form was lost,and the sparse was scattered,the density was decreased,wall capacity decreased,the number of viability decreased,cell wall shrinkage,cytoplasm concentration,light transmittance decreased,the cells from the normal colony growth becomes larger and larger,the connection between the reduction,resulting in apoptotic bodies,Rhizoma Amorphophalli of ethyl acetate extract could significantly induce tumor cell apoptosis;Flow cytometry showed that the apoptotic rate increased significantly with the increase of the concentration and time of the ethyl acetate extract,indicating that the ethyl acetate site had better activity to inhibit the proliferation of tumor cells.Conclusion: 1.The traditional identification method combined with DNA bar code molecular identification technology to determine the Rhizoma Amorphophalli of medicinal plants the original,comprehensively enhance the quality of herbal medicine evaluation standards,the establishment of a quick and better distinction between the Rhizoma Amorphophalli genuine and falsify.2.The anti-tumor activity and active site of Rhizoma Amorphophalli in vitro were determined by CCK-8 method.The inhibitory rate and the IC50 value of the four kinds of tumor cells were obtained.The inhibition rate of the ethyl acetate was higher than that of the other polar parts of the She liu gu,and the IC50 of the human ovarian cancer cells Thelowest value;Cell morphology observation and flow cytometry showed that the activity of ethyl acetate was stable to the tumor cells,and laid a foundation for the further study of anti-tumor active components and anti-tumor mechanism.